构建具有增强抗HIV活性的蓝藻抗病毒蛋白N的强制结构域交换二聚体。
Engineering an obligate domain-swapped dimer of cyanovirin-N with enhanced anti-HIV activity.
作者信息
Kelley Brendan S, Chang Leng Chee, Bewley Carole A
机构信息
Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, MD 20892-0820, USA.
出版信息
J Am Chem Soc. 2002 Apr 3;124(13):3210-1. doi: 10.1021/ja025537m.
The anti-HIV cyanobacterial protein cyanovirin-N can undergo domain swapping to form an intertwined dimer. The dimeric form is stable at low pH and millimolar concentrations. By deleting an amino acid from the hinge linker about which domain swapping occurs, we have constructed an obligate domain-swapped dimer of cyanovirin-N that represents a new tetravalent carbohydrate binding protein that is stable over a large range of pH values. This obligate dimer displays enhanced anti-HIV activity relative to the wild-type cyanovirin-N monomer with an observed 3.5-fold decrease in IC(50) (9nM for the dimer vs 32 nM for the monomer) for inhibition of HIV-1 envelope-mediated cell fusion and, when expressed in Escherichia coli, can be rapidly obtained in >98% purity in a single chromatographic step.
抗HIV蓝藻蛋白氰病毒素-N可发生结构域交换形成交织的二聚体。二聚体形式在低pH值和毫摩尔浓度下稳定。通过从发生结构域交换的铰链连接区删除一个氨基酸，我们构建了一种专一性结构域交换的氰病毒素-N二聚体，它代表一种新的四价碳水化合物结合蛋白，在很大的pH值范围内都很稳定。相对于野生型氰病毒素-N单体，这种专一性二聚体表现出增强的抗HIV活性，在抑制HIV-1包膜介导的细胞融合方面，观察到IC(50)降低了3.5倍（二聚体为9 nM，单体为32 nM），并且在大肠杆菌中表达时，可通过单一色谱步骤快速获得纯度>98%的产物。
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